Long-term exposure to waterborne nonylphenol alters reproductive physiological parameters in economically important marine fish.

Low concentrations of nonylphenol (NP) in aquatic environment can induce drastic effects on the endocrine system in animals. In this study, we examined the modulatory effects of NP on reproductive and physiological parameters in juveniles of the red seabream and black rockfish following waterborne NP exposure (0, 1, 10, and 50 µg L-1) for 60 days. In red seabream exposed to 50 µg L-1 NP, plasma levels of 17ß-estradiol (E2) and 11-ketotestosterone (11-KT) were significantly lower at 30 and 60 days, while E2 levels were slightly higher in 10 µg L-1-exposed individuals at day 30. Similarly, significantly lower levels of E2 and 11-KT were observed in 10 and 50 µg L-1-exposed black rockfish at 60 days, whereas the E2 level was higher in 1 µg L-1-exposed individuals at day 30. After exposure to NP, plasma and mRNA levels of vitellogenin (VTG) were significantly higher in both species at 30 and 60 days, similar to the inducible effects from synthetic estrogen. Plasma cortisol levels were significantly elevated by relatively higher concentrations of NP (10 and 50 µg L-1) at 30 and 60 days. Finally, 60 days of exposure of 50 µg L-1 NP significantly decreased the gonadosomatic index (GSI) and increased the hepatosomatic index (HSI) in both species. The results obtained from this study provide an evidence of the endocrine disrupting potential of waterborne NP on early stages of economically important marine fish. The NP-triggered endocrine modulation can induce effects on the development of reproductive and metabolic organs in fish species.

Haematological assessment of four amazonian ornamental armoured catfish (Teleostei, Loricariidae) / Avaliação hematológica em quatro acaris ornamentais amazônicos (Teleostei, Loricariidae)

The aim of this study was to characterize the haematological profile of four Amazonian ornamentalfreshwater armoured catfish: acari-bola (Peckoltia oligospila - L06), acari-pleco (Cochliodon sp. - L145), acari-canoa(Lasiancistrus saetiger - L323) and acari-assacu (Pseudocanthicus spinosus - L160). The highest blood glucose levels(72.47 ± 28.7 mg dL-1) and erythrocyte counts (0.51 ± 0.2 x 106 cel. μL-1) were recorded for acari-canoa. Theacari-bola and acari-pleco presented similar concentrations of total plasma protein (TPP) (7.96 ± 1.8 and 7.93 ±1.8 g dL-1, respectively) against lower TPP concentrations observed in acari-canoa (4.87 ± 1.5 g dL-1) and acariassacu(6.55 ± 1.5 g dL-1). The acari-assacu had lower total haemoglobin concentration (5.88 ± 1.7 g dL-1) andhaematocrit (12.66 ± 4.6%). No interspecific differences were observed in mean corpuscular volume (MCV) andmean corpuscular haemoglobin concentration (MCHC). The acari-pleco and acari-assacu presented the highestcounts for neutrophils (4142.42 ± 3280.1 cel. μL-1) and thrombocytes (4778.33 ± 1224.8 cel. μL-1), respectively.The haematological profiles were similar to those reported in the literature for freshwater fish and the interspecificdifferences observed were discussed.

O objetivo deste estudo foi caracterizar os perfis hematológicos de quatro espécies de acarisornamentais da região amazônica: acari-bola (Peckoltia oligospila - L06), acari-pleco (Cochliodon sp. - L145), acaricanoa(Lasiancistrus saetiger - L323) e acari-assacu (Pseudocanthicus spinosus - L160). Os maiores níveis de glicose(72,47 ± 28,7 mg dL-1) e contagem de eritrócitos (0,51 ± 0,2 x 106 cel. μL-1) foram encontrados no acari-canoa. Oacari-bola e acari-pleco apresentaram concentrações semelhantes de proteína plasmática total (PPT) (7,96 ± 1,8 e7,93 ± 1,8 g dL-1, respectivamente) em oposição às menores concentrações de PPT no acari-canoa (4,87 ±1,5 g dL-1) e acari-assacu (6,55 ± 1,5 g dL-1). Essa última espécie apresentou menor concentração total dehemoglobina (5,88 ± 1,7 g dL-1) e hematócrito (12,66 ± 4,6%). Não foram observadas diferenças significativasentre as quatro espécies nos parâmetros volume corpuscular médio (VCM) e concentração de hemoglobinacorpuscular média (CHCM). O acari-pleco e o acari-assacu apresentaram os maiores valores de neutrófilos(4142,42 ± 3280,1 cel. μL-1) e de trombócitos (4778,33 ± 1224,8 cel. μL-1), respectivamente. Os perfishematológicos foram semelhantes aos relatados na literatura para peixes de água doce e as diferençasinterespecíficas observadas foram discutidas.

Genotoxic and mutagenic assessment of iron oxide (maghemite-γ-Fe2O3) nanoparticle in the guppy Poecilia reticulata.

The environmental risk of nanomaterials (NMs) designed and used in nanoremediation process is of emerging concern, but their ecotoxic effects to aquatic organism remains unclear. In this study, the citrate-coated (maghemite) nanoparticles (IONPs) were synthesized and its genotoxic and mutagenic effects were investigated in the female guppy Poecilia reticulata. Fish were exposed to IONPs at environmentally relevant iron concentration (0.3 mg L-1) during 21 days and the animals were collected at the beginning of the experiment and after 3, 7, 14 and 21 days of exposure. The genotoxicity and mutagenicity were evaluated in terms of DNA damage (comet assay), micronucleus (MN) test and erythrocyte nuclear abnormalities (ENA) frequency. Results showed differential genotoxic and mutagenic effects of IONPs in the P. reticulata according to exposure time. The IONP induced DNA damage in P. reticulata after acute (3 and 7 days) and long-term exposure (14 and 21 days), while the mutagenic effects were observed only after long-term exposure. The DNA damage and the total ENA frequency increase linearly over the exposure time, indicating a higher induction rate of clastogenic and aneugenic effects in P. reticulata erythrocytes after long-term exposure to IONPs. Results indicated that the P. reticulata erythrocytes are target of ecotoxicity of IONPs.

Genotoxic and hematological parameters in Colossoma macropomum (Pisces, Serrasalmidae) as biomarkers for environmental impact assessment in a protected area in northeastern Brazil.

Genotoxic and hematological parameters in tambaqui (Colossoma macropomum), a native freshwater fish, were used as biomarkers to assess exposure to environmental stressors within the Maracanã Protected Area of Maranhão State, Brazil. Fish were sampled at two sites-Serena Lagoon (control) and Ambude River-on four occasions (dry and rainy season), and biometric data (length and weight) recorded and blood collected from all fish for analysis. Erythrocyte indices-mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration-were calculated. Blood samples were stained with Giemsa and acridine orange, and microscopically examined for micronuclei and morphological nuclear abnormalities. Micronuclei were observed in fish from both sites, although the frequency was significantly higher in fish from the Ambude River and morphological nuclear abnormalities were only observed in fish from the Ambude River. More morphological nuclear abnormalities and a larger number of micronuclei were observed in erythrocytes stained with acridine orange compared with those stained with Giemsa. On average, erythrocyte indices were lower in fish from the Ambude River than from the Serena Lagoon. The results confirm that genotoxic and hematological parameters in C. macropomum can be used as indicators of environmental health and could be valuable tools for monitoring environmental conditions within protected areas.

Hematological and morphometric blood value of four cultured species of economically important tropical foodfish

The use and validation of fish health monitoring tools have become increasingly evident due to aquaculture expansion. This study investigated the hematology and blood morphometrics of Piaractus mesopotamicus, Brycon orbignyanus, Oreochromis niloticus and Rhamdia quelen. The fish were kept for 30 days in 300-liter aquariums, after which they were anesthetized with benzocaine and blood was collected from caudal vessels. In comparison to other species, B. orbignyanus presented the highest hematocrit (Ht), RBC averages and Mean Corpuscular Volume (MCV) with a particular range of data. B. orbignyanus presented lower Ht, Hb, RBC averages and values, and Mean Corpuscular Hemoglobin Concentration (MCHC). Oreochromis niloticus presented lower Ht, Hb, RBC averages and values, and Mean Corpuscular Hemoglobin Concentration (MCHC). Rhamdia quelen and O. niloticus presented higher variation of White Blood Cells (WBC), neutrophils (Nf), lymphocytes (Lf), monocytes (Mf) and thrombocytes (Trb). Data of large axes (LA), minor axes (MA), surface (SF) and volume (VL) are in the same variance range. This study has demonstrated that hematological variances can occur between animals of different species as well as of the same species.

O uso e validação de ferramentas para monitorar o estado de saúde dos peixes está se tornando mais evidente, devido à expansão da aquicultura. Este estudo investigou a hematologia e a morfometria do sangue de Piaractus mesopotamicus, Brycon orbignyanus, Oreochromis niloticus e Rhamdia quelen. Os peixes foram mantidos por 30 dias em aquários de 300 litros, após esse período os peixes foram anestesiados com benzocaína e o sangue coletado por punção caudal. Em comparação com outras espécies B. orbignyanus apresentou maiores médias de hematócrito (Ht), eritrócitos (Eri) e volume corpuscular médio (VCM) com um determinado intervalo de dados. Oreochromis niloticus apresentou menor média e valor de Ht, Hb, eritrócitos e concentração de hemoglobina corpuscular média (CHCM). Rhamdia quelen e O. niloticus apresentaram maior variedade de leucócitos (Leu), neutrófilos (Nf), linfócitos (Lf), monócitos (Mf) e de trombócitos (Trb). Dados de eixos maiores (EMA), eixos menores (EME), superfície (SF) e volume (VL) estão no mesmo intervalo de variação. Este estudo demonstrou que as variações hematológicas não ocorrem somente entre os animais de diferentes espécies, mas também entre os indivíduos da mesma espécie.

Biochemical markers for the assessment of pollution of selected small streams in the Czech Republic.

OBJECTIVES: The objective of the presented study was to investigate levels of the most important biochemical markers in fish which were caught upstream and downstream from sites near waste water treatment plants situated at 3 selected small streams of the Czech Republic. Organic pollutants and mercury were determined in muscle tissue of fish for complex assessment. DESIGN: Levels of biochemical markers (cytochrome P450 (CYP450), ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), vitellogenin (VTG)) were measured in selected tissues (liver, plasma, brain, gill and kidney) samples of brown trout. The concentrations of persistent organic pollutants (POPs) and mercury were determined in muscle samples. RESULTS: Significantly higher levels of biochemical markers (EROD, GST, VTG) were measured downstream from waste water treatment plant (WWTP) in Vlachovo Brezí (the small Libotýnský brook). Values of biochemical markers at the other localities Cerný brook and Moravice River were measured downstream from WWTP consistently or slightly lower with values of upstream. The values of POPs and mercury were in all monitored sites higher upstream compared with downstream from WWTP. The highest values of POPs and mercury were found on Cerný brook in Bruntál (more industrial and agricultural activity). CONCLUSION: The highest occurrence of selected POPs and mercury was generally upstream suggesting that small WWTP are able to partially remove these substances in those locations. But finding higher values of biochemical markers in downstream shows that there are many other substances endocrine disrupting chemicals (EDCs) that WWTP are unable to remove.

Assessment of spermatogenesis and plasma sex steroids in a seasonal breeding teleost: a comparative study in an area of influence of a tributary, downstream from a hydroelectric power dam, Brazil.

River damming and building of hydroelectric power plants interrupt the reproductive migration routes and change the major physicochemical parameters of water quality, with drastic consequences for populations of migratory fishes. The goal of this study was to evaluate proliferation and cell death during spermatogenesis and serum profiles of sex steroids in Prochilodus argenteus, from the São Francisco River, downstream from the Três Marias Dam. A total of 257 adult males were caught quarterly during a reproductive cycle in two sites: the first 34 km of the river after the dam (site 1) and the second 34-54 km after the dam (site 2), after the confluence with a tributary, the Abaeté River. Seasonal changes in the testicular activity associated with morphometric analyses of germ cells as well as proliferation and testicular apoptosis support a more active spermatogenesis in fish from site 2, where higher levels of sex steroids and gonadosomatic index (GSI) were also found. In site 1, fish presented low serum levels of testosterone, 17ß-estradiol and 17α-hydroxyprogesterone and a low GSI during gonadal maturation. Spermatogonial proliferation (PCNA) and apoptosis (TUNEL) were more elevated in fish from site 1, but spermatocytes were mainly labelled in fish from site 2. Overall, these data demonstrate changes in testicular activity and plasma sex steroids in a neotropical teleost fish living downstream from a hydroelectric dam, supplying new data on fish reproduction in regulated rivers. Moreover, morphometric analyses associated with sex steroids profiles provide reliable tools to assess fish spermatogenesis under environmental stress conditions.

Prioritising pharmaceuticals for environmental risk assessment: Towards adequate and feasible first-tier selection.

The presence of pharmaceuticals in the aquatic environment, and the concerns for negative effects on aquatic organisms, has gained increasing attention over the last years. As ecotoxicity data are lacking for most active pharmaceutical ingredients (APIs), it is important to identify strategies to prioritise APIs for ecotoxicity testing and environmental monitoring. We have used nine previously proposed prioritisation schemes, both risk- and hazard-based, to rank 582 APIs. The similarities and differences in overall ranking results and input data were compared. Moreover, we analysed how well the methods ranked seven relatively well-studied APIs. It is concluded that the hazard-based methods were more successful in correctly ranking the well-studied APIs, but the fish plasma model, which includes human pharmacological data, also showed a high success rate. The results of the analyses show that the input data availability vary significantly; some data, such as logP, are available for most API while information about environmental concentrations and bioconcentration are still scarce. The results also suggest that the exposure estimates in risk-based methods need to be improved and that the inclusion of effect measures at first-tier prioritisation might underestimate risks. It is proposed that in order to develop an adequate prioritisation scheme, improved data on exposure such as degradation and sewage treatment removal and bioconcentration ability should be further considered. The use of ATC codes may also be useful for the development of a prioritisation scheme that includes the mode of action of pharmaceuticals and, to some extent, mixture effects.

Kinetically-calibrated solid-phase microextraction using label-free standards and its application for pharmaceutical analysis.

Pre-equilibrium solid-phase microextraction (PE-SPME) has attracted considerable research attention due to shorter sampling times and better temporal resolution than afforded by equilibrium SPME (E-SPME). However, the calibration of PE-SPME is often time-consuming and requires deuterated calibrants, which if available, are often expensive. To address these challenges, we propose a simple but versatile kinetic calibration method, in which nonisotopic (label-free) compounds of interest can supplant the use of deuterated analogues, and the need to determine partitioning coefficients inherent to earlier procedures has been eliminated. Using this approach, both free and total concentrations of analytes can be simultaneously measured within complex sample systems with high accuracy and precision. This calibration method was validated against established E-SPME and solid-phase extraction techniques through the measurement of selected pharmaceuticals in progressively complex matrixes including inorganic buffers, fish blood, and municipal wastewater effluents. This calibration approach may significantly improve time and cost-effectiveness, while improving the application of the SPME approach within highly dynamic systems.

An environmental risk assessment for oseltamivir (Tamiflu) for sewage works and surface waters under seasonal-influenza- and pandemic-use conditions.

In the event of an influenza pandemic, anti-viral medications such as oseltamivir (Tamiflu) are expected to be used in high amounts over a duration of several weeks. Oseltamivir has been predicted to reach high concentrations in surface waters and sewage works. New oseltamivir environmental fate and toxicity studies permit an environmental risk assessment (ERA) under seasonal- and pandemic-use scenarios. The environmental fate data for sewage works (no removal), surface waters (no significant degradation), and water/sediment systems (>50% primary degradation in 100 days) were used for the derivation of new predicted environmental concentrations (PECs) for western Europe and the River Lee catchment in the UK. Existing worst-case PECs for western Europe, the River Lee catchment in the UK and the Lower Colorado basin in the USA under pandemic conditions (< or =98.1 microg/L for surface waters, < or =348 microg/L for sewage works) were also considered for the ERA. PECs were compared with predicted no-effect concentrations (PNECs) based on new chronic ecotoxicity data (no observed effect concentration for algae, daphnia, and fish > or =1 mg/L). Based on all PEC/PNEC risk ratios, no significant risk is evident to surface waters or sewage works during both regular seasonal-use and high pandemic-use of oseltamivir.

Assessment of micronuclei induction in peripheral erythrocytes of fish exposed to xenobiotics under controlled conditions.

The aim of the present study was to standardize and to assess the predictive value of the cytogenetic analysis by MN test in fish erythrocytes as a biomarker for marine environmental contamination. MN frequency baseline in erythrocytes was evaluated in a number of fish species from a reference area (S. Teresa, La Spezia Gulf) and genotoxic potential of a number of common chemical contaminants and mixtures was determined in fish experimentally exposed in aquarium under controlled conditions. Fish (Scophthalmus maximus) were exposed for 3 weeks to 50 ppb of single chemicals (dialkyl phthalate, bisphenol A, tetrabromodiphenyl ether), 30 ppb nonylphenol and mixtures (North Sea oil and North Sea oil with alkylated phenols). Chromosomal damage was determined as micronuclei (MN) frequency in fish erythrocytes. Nuclear anomalies such as blebbed, notched and lobed nuclei were also recorded. Significant increase in MN frequency was observed in erythrocytes of fish exposed to bisphenol A and tetrabromodiphenylether. Chemical mixture North Sea oil+alkylated phenols induced the highest MN frequency (2.95 micronucleated cells/1000 cells compared to 1 MNcell/1000 cells in control animals). The study results revealed that micronucleus test, as an index of cumulative exposure, appears to be a sensitive model to evaluate genotoxic compounds in fish under controlled conditions.

An integrated assessment of estrogenic contamination and biological effects in the aquatic environment of The Netherlands.

An extensive study was carried out in the Netherlands on the occurrence of a number of estrogenic compounds in surface water, sediment, biota, wastewater, rainwater and on the associated effects in fish. Compounds investigated included natural and synthetic hormones, phthalates, alkylphenol(ethoxylate)s and bisphenol-A. The results showed that almost all selected (xeno-)estrogens were present at low concentrations in the aquatic environment. Locally, they were found at higher levels. Hormones and nonylphenol(ethoxylate)s were present in concentrations that are reportedly high enough to cause estrogenic effects in fish. Field surveys did not disclose significant estrogenic effects in male flounder (Platichthys flesus) in the open sea and in Dutch estuaries. Minor to moderate estrogenic effects were observed in bream (Abramis brama) in major inland surface waters such as lowland rivers and a harbor area. The prevalence of feminizing effects in male fish is largest in small regional surface waters that are strongly influenced by sources of potential hormone-disrupting compounds. High concentrations of plasma vitellogenin and an increased prevalence of ovotestes occurred in wild male bream in a small river receiving a considerable load of effluent from a large sewage treatment plant. After employing in vitro and in vivo bioassays, both in situ and in the laboratory, we conclude that in this case hormones (especially 17 alpha-ethynylestradiol) and possibly also nonylphenol(ethoxylate)s are primarily responsible for these effects.